猫疱疹病毒1型糖蛋白B（gp143/108）的表达与鉴定

Expression and identification of the feline herpesvirus type 1 glycoprotein B (gp143/108).

作者信息

Maeda K, Hirasawa K, Kawaguchi Y, Ono M, Mori T, Gemma T, Yokoyama N, Doi K, Mikami T

机构信息

Department of Veterinary Microbiology, Faculty of Agriculture, University of Tokyo, Japan.

出版信息

Virus Res. 1995 Nov;39(1):55-61. doi: 10.1016/s0168-1702(95)00059-3.

DOI:10.1016/s0168-1702(95)00059-3

PMID:8607284

Abstract

The gene for feline herpesvirus type 1 (FHV-1) glycoprotein B (gB) has been cloned into an expression vector, pRVSVneo, containing the long terminal repeat of Rous sarcoma virus and polyadenylation signal of SV40. This expression vector containing FHV-1 gB gene, pRVSVgBneo, was transfected into Crandell feline kidney (CRFK) cells which are susceptible to FHV-1 infection. By indirect immunofluorescence analysis, the expressed gB was recognized with a panel of monoclonal antibodies (MAbs) against FHV-1 gp143/108. Immunoprecipitation analysis using a MAb 34H12 showed that molecular weights of the gB were 143 and 108 kDa under non-denaturing conditions that 108, 70, 64, and 58 kDa under denaturing conditions. The molecular weights were similar to those of the gB expressed in FHV-1-infected CRFK cells. In addition, when plasmid DNAs were injected into mice to obtain gB-monospecific serum, the pooled serum from mice inoculated with pRVSVgBneo, but not with pRVSVgDneo or pRVSVneo, recognized the FHV-1 gB polypeptides.

摘要

猫疱疹病毒1型（FHV-1）糖蛋白B（gB）基因已被克隆到一个表达载体pRVSVneo中，该载体含有劳氏肉瘤病毒的长末端重复序列和SV40的聚腺苷酸化信号。这个含有FHV-1 gB基因的表达载体pRVSVgBneo被转染到对FHV-1感染敏感的克兰德尔猫肾（CRFK）细胞中。通过间接免疫荧光分析，表达的gB能被一组抗FHV-1 gp143/108的单克隆抗体（MAb）识别。使用单克隆抗体34H12进行的免疫沉淀分析表明，在非变性条件下gB的分子量为143和108 kDa，在变性条件下为108、70、64和58 kDa。这些分子量与在FHV-1感染的CRFK细胞中表达的gB的分子量相似。此外，当将质粒DNA注射到小鼠体内以获得gB单特异性血清时，接种pRVSVgBneo而非pRVSVgDneo或pRVSVneo的小鼠的混合血清能识别FHV-1 gB多肽。